Serum adipokines and gene polymorphisms in peripheral vascular disease
-
Fatih Yanar
,
Sercan Yuksel
,
Hasan Basoglu
,
Ibrahim Yilmaz Basar
and
Elif Ozkok
Abstract
Objectives
Peripheral vascular disease (PVD) is an atherosclerotic disease by the occlusion of arteries in the lower extremities. Buerger’s disease is autoimmune due to inflammation of small and medium-sized vessels. The aim was to investigate the relations among adiponectin, leptin, resistin, and variants of ADIPOQ -11,377 C/G, LEP2548G/A, RETN -420 G/C in PVD and Buerger’s disease.
Methods
The study group consisted of 151 patients (127 PVD, 24 Buerger’s disease). Adiponectin, leptin, and resistin were analyzed by ELISA. ADIPOQ −11377 C/G, LEP2548G/A, and RETN -420 G/C were genotyped with designed primers, and probes in LightCycler 480.
Results
Adiponectin was found to be insignificantly higher in the PVD than that Buerger’s disease (p=0.17). Leptin was found to be significantly lower in the Buerger’s disease than in the PVD (p=0.018). The relationships were among age and leptin (r= −0.264; p=0.047), ankle brachial index (ABI) (r= −0.206; p=0.045), adiponectin (r=0.210; p=0.026) in PVD. In Buerger’s disease, resistin and leptin had a significant relationship (r= −0.508; p=0.019).
Conclusions
It is thought that the high serum leptin levels observed in the Buerger group carrying the ADIPOQ GG genotype and the PVD group carrying the RETN GG genotype may be related to the etiopathogenesis of both diseases. Our study is the first to examine the proteins and genetic variations of adipokines in the PVD and Buerger’s disease in Turkish population, and will provide direction for future studies to be conducted with the variants of leptin, adiponectin, resistin, and related proteins.
Introduction
Peripheral vascular disease (PVD) is a progressive atherosclerotic disease characterized by occlusion of arteries in the lower extremities [1], 2]. Inflammation is important for the initiation and progression of PVD.
Buerger’s disease or thromboangiitis obliterans (TAO) is a disorder characterized by inflammation of small and medium-sized arteries and veins in the upper and lower extremities. It is a non-atherosclerotic disease and accounts for 40–60 % of peripheral vascular diseases [3].
Adiponectin has anti-inflammatory and anti-atherogenic effects. Low serum adiponectin levels are an independent risk factor for endothelial dysfunction and are associated with cardiovascular disease [4]. Symptomatic atherosclerotic peripheral arterial disease (PAD) has been associated with decreased adiponectin levels and impaired vascular reactivity [5]. One of the variants identified in the regulatory region of the adiponectin gene (ADIPOQ), the −11377 C/G variant, has been shown to affect serum adiponectin and is associated with risk factors such as hypertension, dyslipidemia, obesity, and type 2 diabetes mellitus (T2DM) [6], [7], [8], [9]. The rs266729 and rs182052 SNPs in the ADIPOQ gene are associated with adiponectin levels, and the minor alleles of these regions have been associated with the prevalence of coronary artery disease (CAD) and cardiovascular and metabolic diseases, including unstable angina and T2DM [9], [10], [11].
Leptin is a pro-inflammatory adipokine that stimulates vascular inflammation and causes vascular smooth muscle hypertrophy, which may contribute to the pathogenesis of T2DM, hypertension, atherosclerosis, and coronary heart disease (CHD) [11]. −2,548 G>A (rs7799039) is a common variant of the leptin promoter, resulting in a guanine to adenine change at −2,548 upstream of the ATG start site [12]. Furthermore, the −2,548 G>A LEP polymorphism has been reported to have a strong role in the transcriptional activation of the LEP promoter by insulin-mediated glucose metabolism [13].
The Resistin (RETN) -420 G>C rs1862513 polymorphism in the 5′ promoter region of the resistin-encoding gene (RETN) has been associated with cardiovascular risk factors [14]. In particular, the -420 C>G polymorphism has been reported to have a potential role in vascular inflammation and the development and progression of abdominal aort aneurysm (AAA) [15].
Our study aimed to investigate relationship between serum adipokine (adiponectin, leptin and resistin) levels and their gene variants ADIPOQ -11,377 C/G rs266729, LEP2548G/A rs7799039 and RETN -420 G/C rs1862513 polymorphisms with PVD and Buerger’s disease.
Materials and methods
Subject selection
The study group consisted of 151 patients with 127 PVD (female/male: 35/92) and 24 with Buerger’s disease (female/male: 1/23). All patients were admitted to the Peripheral Vascular Surgery Unit, Istanbul University, Istanbul Medical Faculty. Our study was conducted with the approval of Istanbul University Istanbul Medical Faculty Clinical Research Ethics Committee (2015/615).
The ages of PVD were between 60 and 82 years (71.6 ± 11), and Buerger’s disease were between 49 and 66 years (57.6 ± 8.6). Aneurysmatic diseases, Behcet’s disease, connective tissue disease, and chronic renal disease were considered as exclusion criteria for both groups. The atherosclerotic patient group with below-knee involvement was accepted as inclusion criteria for PVD. For Buerger’s group, patients with segmental inflammatory occlusive disease in the lower and upper extremities, frequently affecting medium-sized muscles and small-sized arteries and veins of the extremities, were included in the group.
Ankle-brachial index (ABI) was estimated by measuring the systolic blood pressure at the ankles and arms using a hand-held Doppler, and then calculating a ratio in patients with both PVD and Buerger’s disease. In our study, those with an ABI classification below 0.6 were considered serious or severely pathological, those with an ABI classification between 0.9 and 0.6 were considered moderately pathological, and those with an ABI greater than 0.9 were considered normal [16]. ABI index less than 0.6 was taken as borderline diagnostic criterion for PVD.
A positive history of hypertension was defined as the need for antihypertensive medications or systolic blood pressure of 160 mmHg or greater, or diastolic blood pressure of 95 mmHg or greater. A positive history of hypercholesterolemia was defined as the need for cholesterol-lowering medications or serum cholesterol concentrations of 5.0 mmol/L or above. Cardiovascular risk factors (diabetes mellitus, hypertension, dyslipidemia, smoking) were also evaluated as clinical features in patients with PVD and Buerger’s disease.
The operations and applications that PVD and Buerger’s disease underwent together with their accompanying diseases are shown in Table 1. After surgery, 100 mg/day salicylate was started and general health examinations were performed at 6-month intervals. Any patient who had a smoking habit in the last 10 years was defined as a smoker.
Demographic, clinical, and biochemical evaluations of PVD and Buerger’s disease groups.
| All patient groups (n=151) | PVD (n=127) | Buerger’s disease (n=24) | p-Valueb 95 % CI |
|
|---|---|---|---|---|
| Gender (male/female) (% male) |
(115/36) (76.1) |
(92/35) (72.4) |
(23/1) (95.8) |
0.014 0.66–0.86 |
| Age, yeara | 69.2 ± 11.7 | 71.6 ± 11 | 57.6 ± 8.6 | <0.001 8.9–18.6 |
| Diabetes mellitus, n (%) | 102 (70.83) | 98 (81) | 4 (17.4) | <0.001 1.90–11.39 |
| Hypertension, n (%) | 74 (52.48) | 69 (58.5) | 5 (21.7) | 0.001 1.22–5.93 |
| Hyperlipidemia, n (%) | 14 (11.57) | 13 (13.0) | 1 (4.8) | 0.256 0.37–19.75 |
| Coronary heart disease, n (%) | 45 (32) | 42 (35.6) | 3 (14.3) | 0.108 1.03–8.66 |
| Passed cerebrovascular disease n (%) | 8 (6.89) | 8 (8.2) | – | 0.212 0.86–0.97 |
| Smoking, % Smoker Non-smoker Quit smoker |
31.25 54.86 13.88 |
27.5 58.3 14.2 |
52.2 34.8 13.0 |
0.053 0.34–0.83 |
| ABIa | 0.582 ± 0.36 | 0.62 ± 0.03 | 0.32 ± 0.08 | 0.006 0.087–0.49 |
| Applied wound care/ amputation/debridement procedure, n (%) Resting pain Wound cleansing/debridement Finger or small joint amputation Major joint amputation Trans metatarsal amputation Graft application Bypass Angioplasty |
11 (8.33) 33 (25) 43 (32.57) 21 (15.90) 9 (6.81) 4 (3.03) 7 (5.30) 4 (3.03) |
11 (9.9) 30 (27) 31 (27.9) 18 (16.2) 6 (5.4) 4 (3.6) 7 (6.3) 4 (3.69) |
0 (0) 3 (14.3) 12 (57.1) 3 (14.3) 3 (14.3) 0 (0) 0 (0) 0 (0) |
|
| hs-CRPa, mg/L | 1.52 ± 0.71 | 1.62 ± 0.76 | 1.29 ± 0.55 | 0.09 (−0.14)–0.78 |
| Adiponectina, ng/mL | 4.05 ± 0.35 | 4.07 ± 0.37 | 3.96 ± 0.29 | 0.17 (−0.047)–0.26 |
| Resistina, pg/mL | 4.08 ± 0.26 | 4.08 ± 0.26 | 4.08 ± 0.27 | 0.90 (−0.14)–0.12 |
| Leptina, pg/mL | 0.71 ± 0.65 | 0.82 ± 0.62 | 0.41 ± 0.64 | 0.018 0.067–0.74 |
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a, mean ± SD; CI, confidence interval; SD, standard deviation; ABI, ankle brachial index; PVD, peripheral vascular disease; hs-CRP, high-sensitive C-reactive protein; b, PVD and Buerger’s disease groups were compared. Numbers in bold indicate statistically significant values.
Written informed consent was taken from all subjects for the study.
Biochemical determination
After 12 h of fasting, venous blood samples were collected between 08:00 and 09:00 a.m. into heparin-free vacuum tubes and centrifuged at 400 g for 20 min at ambient temperature to obtain serum samples. Separated serum samples were aliquoted and stored at −20 °C until adipokine analyses. Serum hs-CRP levels were determined immediately.
Determinations of serum adipokines
Serum adipokines were determined by the sandwich enzyme-linked immunosorbent assay (ELISA) technique using plates coated with antibodies specific for human leptin, human adiponectin, and human resistin (Elabscience, USA). A biotinylated detection antibody specific for human adipokines and avidin-horseradish peroxidase (HRP) conjugate were added to each well. Optical density (OD) was measured spectrophotometrically at 450 nm in proportion to the concentrations of adipokines.
The sensitivity reported for serum adiponectin in the kit was 0.10 ng/mL and the detection range was 0.16–10 ng/mL [17]. The sensitivity and detection range of the kit used for serum resistin were 18.75 pg/mL and 31.25–2,000 pg/mL, respectively [18], and the sensitivity and detection range of the kit used for serum leptin were 9.38 pg/mL and 15.63–1,000 pg/mL, respectively [19]. The % coefficient of variation (%CV) for each adipokine is<10 %.
Genotypings of ADIPOQ-11377 C/G, LEP2548 G/A, RETN -420 G/C
Primers and probes designed against gene regions for ADIPOQ -11377 C/G rs266729, LEP2548 G/A rs7799039, and RETN -420 G/C rs1862513 in previously isolated DNA samples were genotyped in LightCycler 480 (Roche, Germany) [20], [21], [22].
Statistical analyses
In our study, parameters showing normal distribution from continuous data were evaluated with the student’ t-test using the SPSS version 21 package program. In this study, hs-CRP, adiponectin, resistin, and leptin results, which did not show normal distribution in our groups, were converted to log10 transformation values. The chi-square test was used to evaluate categorical data. In the PVD and Buerger’s disease, the distribution of adipokine genes within the group was analyzed to see if it was consistent with Hardy-Weinberg [23]. Pearson’s correlation test was applied to evaluate statistical serum protein levels in each group. The Mann-Whitney U test was applied to evaluate the significance of parameters that did not show normal distribution. Using the G power 3.1.9.7 program, when the effect size was entered as 0.7 and the allocation ratio N2/N1: 0.1889, we obtained the power of our study as 0.917. p<0.05 was taken as the significance limit.
Results
Table 1 shows the demographic and clinical findings for all patients and patient groups with PVD and Buerger’s disease. PVD patients (age range: 60.6–82.6) were found to be significantly older than the Buerger’s disease (age range: 49.1–66.3) (p<0.001, 95 % confidence interval (CI): 8.9–18.6). When gender distribution between groups was examined, male gender was found to be higher in the Buerger’s disease (p=0.014, 95 % CI: 0.66–0.86). It was found that smokers were higher in Buerger’s disease, but it was not significant (p=0.053, 95 % CI: 0.34–0.83).
In PVD, 81 % had DM (p<0.001, 95 % CI: 1.90–11.39), and 58.5 % had hypertension (p=0.001, 95 % CI: 1.22–5.93), which was significantly higher than Buerger’s disease. There were no significant differences in hyperlipidemia, coronary heart disease, or passed cerebrovascular disease between groups (p=0.256, 95 % CI: 0.37–19.75; p=0.108, 95 % CI: 1.03–8.66; and p=0.212, 95 % CI: 0.86–0.97, respectively) (Table 1).
When we examined the ABI values of two patient groups, they were found to be significantly higher in the PVD (p=0.006, 95 % CI: 0.087–0.49) (Table 1).
It was found that hs-CRP between groups was insignificantly higher in PVD (p=0.09, 95 % CI: (−0.14)–0.78) (Table 1). The levels of adiponectin were higher in PVD, but did not reach significance (p=0.17, 95 % CI: (−0.047)–0.26); there was also no significant difference between groups in terms of resistin values (p=0.90, 95 % CI: (−0.14)–0.12). Leptin levels were significantly lower in Buerger’s disease than in PVD (p=0.018, 95 % CI: 0.067–0.74) (Table 1).
When we compared hs-CRP and serum adipokine between groups with ABI values below 0.6, we found that hs-CRP, leptin, and adiponectin were higher in PVD (p=0.105, 95 % CI: (−0.18)–1.45; p=0.138, 95 % CI: (−0.13)–0.81; p=0.278, 95 % CI: (−0.069)–0.31, respectively). Resistin was found to be similar between groups with ABI values below 0.6 (p=0.71, 95 % CI: (−0.23)–0.12) (Table 2).
Demographic, clinical, and biochemical evaluations of PVD and Buerger’s disease groups according to ABI classification.
| PVD | Buerger’s disease | |||||
|---|---|---|---|---|---|---|
| ABI<0.6 | ABI=0.6–0.9 | ABI>0.9 | ABI<0.6 | ABI=0.6–0.9 | ABI>0.9 | |
| n (%) | 56 (57.7) | 24 (24.7) | 17 (17.5) | 11 (84.6) | 1 (7.7) | 1 (7.7) |
| Gender (male/Female) (% male) |
(47/9) (83.9) |
(12/12) (50) |
(12/5) (70.6) |
(10/1) (90.9) |
(1/0) (100) |
(1/0) (100) |
| Diabetes mellitus n (%) | 44 (80.0) | 20 (83.3) | 16 (94.1) | 2 (18.2)b | 0 (0) | 0 (0) |
| Hypertension n (%) | 32 (61.5) | 14 (58.3) | 8 (47.1) | 3 (27.3)c | 0 (0) | 1 (100) |
| Hyperlipidemia n (%) | 6 (13.3) | 2 (9.2) | 1 (7.7) | 1 (100) | 0 (0) | 0 (0) |
| Passed cerebrovascular disease n (%) | 5 (11.6) | 3 (13.6) | – | 0 (0) | 0 (0) | – |
| Coronary heart disease n (%) | 21 (39.6) | 9 (37.5) | 5 (31.2) | 1 (10) | 0 (0) | 1 (100) |
| Applied wound care/amputation/debridement procedure n (%) Resting pain Wound cleansing/debridement Finger or small joint amputation Major joint amputation Trans metatarsal amputation Graft application Bypass application |
4 (66.7) 13 (56.5) 11 (42.3) 12 (85.7) 3 (50) 2 (50) 5 (83.3) |
2 (33.3) 3 (13) 9 (34.6) 1 (7.1) 2 (33.3) 1 (25) 1 (16.7) |
– 7 (30.4) 6 (23.1) 1 (7.1) 1 (16.7) 1 (25) – |
– 1 (50) 3 (75) – – 2 (50) – |
– – 1 (25) – – 1 (25) – |
– 1 (50) – 3 (100) 2 (100) 1 (25) – |
| hs-CRPa, mg/L | 1.69 ± 0.72 | 1.76 ± 0.50 | 2.17 ± 0.23 | 1.05 ± 0.68 | 1.09 (1) | – |
| Adiponectina, ng/mL | 4.04 ± 0.35 | 4.02 ± 0.51 | 4.09 ± 0.26 | 3.91 ± 0.25 | 4.38 (1) | 3.95 (1) |
| Resistina, pg/mL | 4.07 ± 0.31 | 4.11 ± 0.19 | 4.04 ± 0.36 | 4.13 ± 0.21 | 0.19 (1) | 3.77 (1) |
| Leptina, pg/mL | 0.67 ± 0.65 | 1.04 ± 0.59 | 0.74 ± 0.58 | 0.33 ± 0.63 | 0.20 (1) | 0.49 (1) |
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a, mean ± SD; SD, standard deviation; ABI, ankle brachial index; CI, confidence interval; hs-CRP, high-sensitive C-reactive protein; PVD, peripheral vascular disease. cp<0.05, versus PVD patients with ABI less than 0.6, 95 % CI: 0.84–6.064; bp<0.001, versus PVD patients with ABI less than 0.6, 95 % CI: 1.24–15.52. ABI classification was made reference to the Ostergren et al. (Reference [16] of this study).
There was a significant difference in adiponectin between the groups in terms of the presence of diabetes (p=0.048, 95 % CI: 0.006–0.75), and hs-CRP was found higher in PVD (p=0.052, 95 % CI: (−0.015)–1.49). There were no differences in resistin and leptin between groups (p=0.73, 95 % CI: (−0.34)–0.24; p=0.069, 95 % CI: (−1.04)–26.65, respectively) (Figure 1).
The levels of hs-CRP, adiponectin, leptin, and resistin in PVD and Buerger’s disease groups with diabetes mellitus. *: p=0.048.
When we selected hypertensive patients in PVD and Buerger’s disease, leptin was found to be significantly higher in PVD (p=0.017, 95 % CI: 0.13–1.31). Resistin and adiponectin were similar in the hypertensive PVD and Buerger’s disease (p=0.86, 95 % CI: (−0.29)–0.24; p=0.81, 95 % CI: (−0.12)–0.16, respectively) (Figure 2).
The levels of hs-CRP, adiponectin, leptin, and resistin in PVD and Buerger’s disease groups with hypertension. *: p=0.017.
Among those with an ABI value less than 0.6, it was observed that those with diabetes were 80 % in the PVD and 18.2 % in the Buerger’s disease (p<0.001, 95 % CI: 1.24–15.52). A significant difference was found in the proportion of hypertensive patients with an ABI value below 0.6, 61.5 % in PVD, and 27.3 % in Buerger’s disease (p=0.038, 95 % CI: 0.84–6.064) (Table 2).
Leptin G2548A rs77999039
LEP G2548A genotype and allele distribution frequencies were found to be similar in groups. The distributions of LEP G2548A within the groups were found to be in accordance with Hardy-Weinberg (for PVD: X2=0.579, p=0.446 and for Buerger’s: X2=0.211, p=0.64, respectively) (Table 3).
Genotype and allele distributions of ADIPOQ, RETN, and LEP in PVD and Buerger’s disease groups.
| PVD, n (%) | Buerger’s disease, n (%) | p-Value | |
|---|---|---|---|
| ADIPOQ -11377 C/G rs266729 CC CG GG C G |
58 (49.6) 49 (41.9) 10 (8.5) 165 (70.5) 69 (29.5) X2=0.0059 p=0.938 |
9 (45) 9 (45) 2 (10) 27 (67.5) 13 (32.5) X2=0.0131 p=0.908 |
NS NS |
| RETN -420 G/C rs1862513 GG GC CC G C |
60 (51.3) 41 (35.0) 16 (13.7) 161 (68.8) 73 (31.2) X2=3.948 p=0.0469 |
7 (36.8) 9 (47.4) 3 (15.8) 23 (60.5) 15 (39.5) X2=0.00143 p=0.969 |
NS NS |
| LEP G2548A rs77999039 GG GA AA G A |
24 (20.7) 53 (45.7) 39 (33.6) 101 (43.5) 131 (56.5) X2=0.579 p=0.446 |
5 (25) 11 (55) 4 (20) 21 (52.5) 19 (47.5) X2=0.211 p=0.64 |
NS NS |
-
NS, non-significant; PVD, peripheral vascular disease. p<0.05 was accepted as a significance value. Courtlab -HW calculator was used for compliance with X2 Hardy-Weinberg Equilibrium. (Reference [23] of this study).
RETN -420 G/C rs1862513
When the RETN -420 G/C genotype and allele distributions were examined, the GG, GC, CC genotypes, and G, C alleles were observed to be different in PVD and the Buerger’s disease, but they were not significant. It was observed that the Hardy-Weinberg distribution was not appropriate within the groups, with X2=3.948, p=0.0469 in PVD; while it conformed to the Hardy-Weinberg with X2=0.00143, p=0.969 in Buerger’s disease (Table 3).
ADIPOQ -11,377 C/G rs266729
Genotype distributions were detected as CC, CG, and GG (for PVD: 49.6 %, 41.9 %, 8.5 %, respectively) (for Buerger’s disease: 45 %, 45 %, 10 %, respectively) in both groups; there were no significant differences. It was found to be similar distributions of C, G allele frequencies in PVD (70.5 %, 29.5 %, respectively) and Buerger’s disease (67.5 %, 32.5 %, respectively). It was determined that both groups conformed to the Hardy-Weinberg, with X2=0.0059, p=0.938 in the PVD, and X2=0.0131 and p=0.908 in Buerger’s disease (Table 3).
A significant relationship was detected between age and continuous variables in PVD, a negative relationship with leptin (r= −0.264, p=0.047), and ABI (r= −0.206, p=0.045), and a positive relationship with adiponectin (r=0.210, p=0.026). There was a significant inverse relationship between resistin and leptin (r= −0.508, p=0.019) in Buerger’s.
According to ADIPOQ -11,377 C/G genotype distributions, leptin was found to be significantly lower in PVD carrying GG (p=0.019, 95 % CI: 2.40–18.55) (Figure 3). Serum leptin was also found to be significantly higher in PVD compared to the Buerger’s disease in those with the RETN -420 GG genotype (p=0.030, 95 % CI: 0.078–1.307) (Figure 4), and there was a limit of significance in those with the LEP2548 GA genotype (p=0.054, 95 % CI: (−0.010)–1.056) (Figure 5). Serum adiponectin was higher in those carrying the LEP2548 AA genotype in the PVD, but were not found to be significant (p=0.063, 95 % CI: (−0.026)–0.93) (Figure 5).
The levels of hs-CRP and adipokines in patients with PVD and Buerger’s disease according to ADIPOQ -11377 C/G rs266729 genotype distributions. *: p=0.019.
The levels of hs-CRP and adipokines in patients with PVD and Buerger’s disease according to RETN -420 G/C rs1862513 genotype distributions. *: p=0.030.
The levels of hs-CRP and adipokines in patients with PVD and Buerger’s disease according to LEP2548 G/A rs7799039 genotype distributions.
Discussion
According to the main findings, leptin in PVD were found to be significantly higher than the values in Buerger’s disease. Adiponectin and leptin were found to be higher in diabetic and hypertensive PVD patients. Among those with ABI values below 0.6, 80 % of diabetic patients were in PVD, and 61.5 % of hypertensive patients were in PVD. It was observed that leptin was significantly lower in PVD carrying the ADIPOQ -11,377 GG, while leptin in those carrying the RETN -420 GG wild-type genotype formed a significant difference.
When we evaluated the age ranges and gender distributions of PVD and Buerger’s disease, we found that the average age was younger in Buerger’s disease, and the male gender was lower in PVD. Buerger’s disease is more common in men than in women, and young and middle-aged men are more affected in the population; the disease rate is three times higher in men than in women [24].
In our study, we found that smoking was higher in the Burger’s disease and that the difference was close to the significance limit in the comparison between the groups. The presence of a smoking history in the pathophysiology of Buerger’s disease has been associated with the incidence and progression in male patients [25].
In this study, we found that hs-CRP was higher in PVD, but it was found to be insignificant. The role of hs-CRP in linking fibrosis and inflammation in the pathophysiology of PVD was demonstrated in patients with PAD studied in the ARIC study [26].
Leptin levels were significantly lower in Buerger’s disease than in PVD, while the decrease in adiponectin levels did not reach a significant level. It was found that serum resistin levels were similar in both PVD and Buerger’s disease.
In this study, depending on the presence of DM, adiponectin and leptin levels were found to be lower in the Buerger’s group than in PVD. Low adiponectin is linked to vascular endothelial dysfunction [5], [6], [7]. Low adiponectin in our Buerger’s disease group may be related to the development of inflammatory reactions [27]. Our study also found that serum leptin was positively associated with PVD in hypertensive patients. There are studies indicating that the leptin system is related to the development of atherosclerosis, which are shown as risk factors in vascular diseases, including PVD and Buerger’s.
Resistin, in contrast, has emerged as a significant player in vascular inflammation and dysfunction. It is secreted by adipocytes and inflammatory cells, particularly in the context of obesity [28]. Resistin has been shown to promote endothelial cell activation, leading to increased expression of adhesion molecules and pro-inflammatory cytokines, which can contribute to atherosclerosis and vascular remodeling [29]. Elevated levels of resistin have been linked to increased risk of cardiovascular events, including ischemic stroke and CAD [30], 31]. Furthermore, resistin’s role in promoting vascular smooth muscle cell migration and proliferation underscores its contribution to the pathogenesis of PVD [32], 33]. In our study, it was found similar resistin values between the PVD and Buerger’s disease groups.
The relationship between these adipokines and PVD is complex and multifaceted. While adiponectin generally exerts protective effects, leptin and resistin are often associated with adverse vascular outcomes. For example, resistin has been implicated in the development of endothelial dysfunction and hypertension, which are critical components of PVD [34], 35]. Studies have demonstrated that high serum resistin levels correlate with increased severity of CAD and PAD, suggesting that resistin may serve as a potential biomarker for vascular damage [36], 37].
It has been shown that an amino acid change occurring in the ADIPOQ gene at the promoter position 11,377 (-11,377 C/G), where cytosine is replaced with guanine, is associated with T2DM. In addition, the G allele of the ADIPOQ rs266729 may appear important in associations with T2DM risk in various populations [38]. Resistin gene is located on chromosome 19p13.2 and spans 1,369 bp with four exons and three introns [39]. RETN -420 C/G in the promoter region of the resistin gene has been identified as the main determinant of plasma resistin concentration and expression in humans [40], 41].
In the PVD group, while leptin was significantly negatively related to age, adiponectin was positively related to age. In studies conducted, higher serum adiponectin in older men and women may be associated with an age-related increase in visceral adipose tissue [42]. The age-related alteration in leptin levels is attributable to changes in fat mass in women and probably also in men [43]. In in vitro studies, Wabitsch et al. [44] demonstrated a direct long-term inhibitory effect of testosterone on leptin production from human adipocytes in culture. It can be explained that the age-related decrease in leptin in the PVD group may be due to the decrease in subcutaneous fat tissue with aging [45].
There was a significant and inverse relationship between age and ABI values in PVD. In a cross-sectional study conducted in Scotland involving 28,980 men and women over the age of 50 without cardiovascular disease, they found that ABI values decreased proportionally with increasing age [46]. In our study, we found that the frequency of diabetics with ABI values lower than 0.6 was significantly higher in PVD. The relationship we found between them can be explained by the fact that the ABI value was reported to be important in the evaluation of diabetic PAD patients, especially in the lower extremities, in cohort studies [47]. We found that patients with hypertension in PVD with an ABI value of less than 0.6 were significantly more likely to have hypertension compared to the Buerger group. In studies, patients with an ABI value of<0.9 were associated with hypertension and a history of acute coronary syndrome were associated with a higher risk of developing heart failure [48].
It was found a difference in serum leptin between PVD and Buerger’s groups carrying RETN wild GG, suggesting that leptin resistance may be more prominent in PVD than in Buerger’s disease, as resistin affects the development of obesity and hyperinsulinemia through glucose and lipid efflux from adipocytes with its physiological effect on leptin. It was observed that the resistin and leptin were significantly inversely correlated in Buerger’s disease. It has been reported that acute resistin treatment inhibited leptin-mediated STAT3 phosphorylation [49]. The inverse relationship between resistin and leptin levels obtained in Buerger’s disease may be due to the central metabolic effect of resistin on leptin, known as leptin resistance, from the finding by Asterholm et al. [50].
The limitation of our study can be shown as the lack of a healthy control group with age and gender-matched with the study groups. The study has an objective cross-sectional study feature for both patient groups. The fact that the study is not a patient-control study eliminates the examination of comparisons with the healthy group. Since there are significant differences between these groups in peripheral vascular diseases, their cross-sectional examination in this way also makes an important contribution to the study.
Conclusions
In conclusion, the roles of adiponectin, leptin, and resistin in both PVD and Buerger’s disease illustrate the intricate balance between protective and harmful effects of adipokines on vascular health. The higher leptin levels we detected in the Buerger group carrying the ADIPOQ GG genotype, and in the PVD group carrying the RETN GG suggest that they may be related to the etiopathogenesis of both diseases. Our study is the first to examine the serum levels and genetic variations of adipokines in PVD and Buerger’s disease in the Turkish population. In addition, these main findings may contribute to the investigations of potential biomarker proteins or genetic variants related to atherosclerosis and inflammation that may be effective in the etiopathogenesis of both PVD and Buerger’s disease.
Funding source: This study was supported by Istanbul University Research Project (Number: TOA-2017-24592)
Award Identifier / Grant number: TOA-2017-24592
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Research ethics: This study was carried out with the approval of Istanbul University Istanbul Medical Faculty Clinical Research Ethics Committee (2015/615).
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Informed consent: Informed consent was obtained from all individuals included in this study.
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Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
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Use of Large Language Models, AI and Machine Learning Tools: None declared.
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Conflict of interest: Authors state no conflict of interest.
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Research funding: This study was supported by Istanbul University Research Project (Project Number: TOA-2017-24592).
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Data availability: All authors confirm that the raw data for the study are made available by the corresponding author when requested.
References
1. Criqui, MH, Matsushita, K, Aboyans, V, Hess, CN, Hicks, CW, Kwan, TW, et al.. American Heart Association Council on Epidemiology and Prevention; council on arteriosclerosis, thrombosis and vascular biology; council on cardiovascular radiology and intervention; council on lifestyle and cardiometabolic health; council on peripheral vascular disease; and stroke council. Lower extremity peripheral artery disease: contemporary epidemiology, management gaps, and future directions: a scientific statement from the American Heart Association. Circulation 2021;144:e171–91. https://doi.org/10.1161/cir.0000000000001005.Search in Google Scholar
2. Signorelli, SS, Marino, E, Scuto, S. Inflammation and peripheral arterial disease. J 2019;2:142–51. https://doi.org/10.3390/j2020012.Search in Google Scholar
3. Fardhani, IM. Buerger’s disease (Thromboangiitis obliterans) among smokers: a literature review. Int J Rare Dis Disord 2023;6:050. https://doi.org/10.23937/2643-4571/1710050.Search in Google Scholar
4. Pasqualini, L, Schillaci, G, Innocente, S, Pucci, G, Coscia, F, Siepi, D, et al.. Lifestyle intervention improves microvascular reactivity and increases serum adiponectin in overweight hypertensive patients. Nutr Metabol Cardiovasc Dis 2010;20:87–92. https://doi.org/10.1016/j.numecd.2009.03.002.Search in Google Scholar PubMed
5. Ho, DY, Cook, NR, Britton, KA, Kim, E, Creager, MA, Ridker, PM, et al.. High-molecular-weight and total adiponectin levels and incident symptomatic peripheral artery disease in women: a prospective investigation. Circulation 2011;124:2303–11. https://doi.org/10.1161/circulationaha.111.045187.Search in Google Scholar
6. Komai, H, Shibata, R, Juri, M, Matsushita, K, Ouchi, N, Murohara, T. Plasma adiponectin as a predictive factor of survival after a bypass operation for peripheral arterial disease. J Vasc Surg 2009;50:95–9. https://doi.org/10.1016/j.jvs.2008.12.044.Search in Google Scholar PubMed
7. Ong, KL, Li, M, Tso, AWK, Xu, A, Cherny, SS, Pak, CS, et al.. Association of genetic variants in the adiponectin gene with adiponectin level and hypertension in Hong Kong Chinese. Eur J Endocrinol 2010;163:251–7. https://doi.org/10.1530/eje-10-0251.Search in Google Scholar PubMed
8. de Luis, DA, Izaola, O, Primo, D, Aller, R. Relation of a variant in adiponectin gene (rs266729) with metabolic syndrome and diabetes mellitus type 2 in adult obese subjects. Eur Rev Med Pharmacol Sci 2020;24:10646–52. https://doi.org/10.26355/eurrev-202010-23422.Search in Google Scholar
9. Smetnev, S, Klimushina, M, Kutsenko, V, Kiseleva, A, Gumanova, N, Kots, A, et al.. Associations of SNPs of the ADIPOQ gene with serum adiponectin levels, unstable angina, and coronary artery disease. Biomolecules 2019;9:537. https://doi.org/10.3390/biom9100537.Search in Google Scholar PubMed PubMed Central
10. Kanu, JS, Qiu, S, Cheng, Y, Li, R, Kou, C, Gu, Y, et al.. Associations between three common single nucleotide polymorphisms (rs266729, rs2241766, and rs1501299) of ADIPOQ and cardiovascular disease: a meta-analysis. Lipids Health Dis 2018;17:126. https://doi.org/10.1186/s12944-018-0767-8.Search in Google Scholar PubMed PubMed Central
11. Singh, M, Bedi, US, Singh, PP, Arora, R, Khosla, S. Leptin and the clinical cardiovascular risk. Int J Cardiol 2010;140:266–71. https://doi.org/10.1016/j.ijcard.2009.07.019.Search in Google Scholar PubMed
12. Gong, DW, Bi, S, Pratley, RE, Weintraub, BD. Genomic structure and promoter analysis of the human obese gene. J Biol Chem 1996;271:3971–4. https://doi.org/10.1074/jbc.271.8.3971.Search in Google Scholar PubMed
13. Bartella, V, Cascio, S, Fiorio, E, Auriemma, A, Russo, A, Surmacz, E. Insulin-dependent leptin expression in breast cancer cells. Cancer Res 2008;68:4919–27. https://doi.org/10.1158/0008-5472.can-08-0642.Search in Google Scholar PubMed
14. Musialik, K, Miller-Kasprzak, E, Walczak, M, Markuszewski, L, Bogdański, P. The association between serum resistin level, resistin (-420C/G) single nucleotide variant, and markers of endothelial dysfunction, including salt taste preference in hypertensive patients. Nutrients 2022;14:1789. https://doi.org/10.3390/nu14091789.Search in Google Scholar PubMed PubMed Central
15. Wu, Z, Liu, S, Yang, Z, Meng, X, An, Y, Zhang, H. Study on the correlation between gene polymorphisms of adiponectin and resistin levels and abdominal aortic aneurysm. Clinics 2023;78:100298. https://doi.org/10.1016/j.clinsp.2023.100298.Search in Google Scholar PubMed PubMed Central
16. Ostergren, J, Sleight, P, Dagenais, G, Danisa, K, Bosch, J, Qilong, Y, et al.. Impact of ramipril in patients with evidence of clinical or subclinical peripheral arterial disease. Eur Heart J 2004;25:17–24. https://doi.org/10.1016/j.ehj.2003.10.033.Search in Google Scholar PubMed
17. Onsun, N, Akaslan, TC, Sallahoglu, K, Gülcan, AS, Bulut, H, Yabacı, A. Effects of TNF inhibitors and an IL12/23 inhibitor on changes in body weight and adipokine levels in psoriasis patients: a 48-week comparative study. J Dermatol Treat 2022;33:1727–32. https://doi.org/10.1080/09546634.2021.1901845.Search in Google Scholar PubMed
18. Hao, Y, Xiao, J, Fu, P, Yan, L, Zhao, X, Wu, X, et al.. Increases in BMI contribute to worsening inflammatory biomarkers related to breast cancer risk in women: a longitudinal study. Breast Cancer Res Treat 2023;202:117–27. https://doi.org/10.1007/s10549-023-07023-w.Search in Google Scholar PubMed
19. Alidu, H, Dapare, PPM, Quaye, L, Amidu, N, Bani, SB, Banyeh, M. Insulin resistance in relation to hypertension and dyslipidaemia among men clinically diagnosed with type 2 diabetes. BioMed Res Int 2023;2023:8873226. https://doi.org/10.1155/2023/8873226.Search in Google Scholar PubMed PubMed Central
20. Herrmann, MG, Durtschi, JD, Bromley, LK, Wittwer, CT, Voelkerding, KV. Amplicon DNA melting analysis for mutation scanning and genotyping: cross-platform comparison of instruments and dyes. Clin Chem 2006;52:494–503. https://doi.org/10.1373/clinchem.2005.063438.Search in Google Scholar PubMed
21. Dujols, V, Kusukawa, N, McKinney, JT, Dobrowolsky, SF, Wittwer, CT. High-resolution melting analysis for scanning and genotyping. In: Tevfik, D, editor. Real-Time PCR. Abingdon: Taylor & Francis; 2006.Search in Google Scholar
22. Reed, GH, Wittwer, CT. Sensitivity and specificity of single-nucleotide polymorphism scanning by high-resolution melting analysis. Clin Chem 2004;50:1748–54. https://doi.org/10.1373/clinchem.2003.029751.Search in Google Scholar PubMed
23. Court, MH, Michael, H. Court’s online calculator. 2005–2008. Court (2005) Court MH. Available from: https://www.scribd.com/doc/246388807/Court-Lab-HW-Calculator.Search in Google Scholar
24. Malecki, R, Zdrojowy, K, Adamiec, R. Thromboangiitis obliterans in the 21st century--a new face of disease. Atherosclerosis 2009;206:328–34. https://doi.org/10.1016/j.atherosclerosis.2009.01.042.Search in Google Scholar PubMed
25. Ketha, SS, Cooper, LT. The role of autoimmunity in thromboangiitis obliterans (Buerger’s disease). Ann N Y Acad Sci 2013;1285:15–25. https://doi.org/10.1111/nyas.12048.Search in Google Scholar PubMed
26. Ding, N, Yang, C, Ballew, SH, Kalbaugh, CA, McEvoy, JW, Salameh, M, et al.. Fibrosis and inflammatory markers and long-term risk of peripheral artery disease: the ARIC study. Arterioscler Thromb Vasc Biol 2020;40:2322–31. https://doi.org/10.1161/atvbaha.120.314824.Search in Google Scholar
27. Ebrahimi-Mamaeghani, M, Mohammadi, S, Arefhosseini, SR, Fallah, P, Bazi, Z. Adiponectin as a potential biomarker of vascular disease. Vasc Health Risk Manag 2015;11:55–70. https://doi.org/10.2147/vhrm.s48753.Search in Google Scholar
28. Scott, T, Babayeva, O, Banerjee, S, Zhong, W, Francis, S. Sgk1 is modulated by resistin in vascular smooth muscle cells and in the aorta following diet‐induced obesity. Obesity 2016;24:678–86. https://doi.org/10.1002/oby.21425.Search in Google Scholar PubMed PubMed Central
29. Jamaluddin, SM, Weakley, SM, Yao, Q, Chen, C. Resistin: functional roles and therapeutic considerations for cardiovascular disease. Br J Pharmacol 2012;165:622–32. https://doi.org/10.1111/j.1476-5381.2011.01369.x.Search in Google Scholar PubMed PubMed Central
30. Rajpathak, S, Kaplan, R, Wassertheil‐Smoller, S, Cushman, M, Rohan, T, McGinn, A, et al.. Resistin, but not adiponectin and leptin, is associated with the risk of ischemic stroke among postmenopausal women: results from the Women’s Health Initiative. Stroke 2011;42:1813–20. https://doi.org/10.1161/strokeaha.110.607853.Search in Google Scholar PubMed PubMed Central
31. Sinan, UY, Canbolat, IP, Baydar, O, Oktay, V, Imre, G, Kocas, C, et al.. Relationship between increased serum resistin level and severity of coronary artery disease. Angiology 2013;65:239–42. https://doi.org/10.1177/0003319713502718.Search in Google Scholar PubMed
32. Ding, Q, Chai, H, Mahmood, N, Tsao, J, Mochly‐Rosen, D, Zhou, W. Matrix metalloproteinases modulated by protein kinase Cε mediate resistin-induced migration of human coronary artery smooth muscle cells. J Vasc Surg 2011;53:1044–51. https://doi.org/10.1016/j.jvs.2010.10.117.Search in Google Scholar PubMed PubMed Central
33. Heun, Y, Gräff, P, Lagara, A, Schelhorn, R, Mettler, R, Pohl, U, et al.. The GEF cytohesin-2/ARNO mediates resistin induced phenotypic switching in vascular smooth muscle cells. Sci Rep 2020;10:3672. https://doi.org/10.1038/s41598-020-60446-z.Search in Google Scholar PubMed PubMed Central
34. Niaz, S, Ijaz, K, Doggar, S, Sheikh, ZI, Zaidi, IA, Irfan, S. Relation of serum resistin with body mass index and lipid profile in hypertensive and ischemic heart disease patients. PJMHS 2021;15:3315–7. https://doi.org/10.53350/pjmhs2115123315.Search in Google Scholar
35. Hsu, BG, Lee, CJ, Yang, CF, Chen, YC, Wang, JH. High serum resistin levels are associated with peripheral artery disease in the hypertensive patients. BMC Cardiovasc Disord 2017;17:80. https://doi.org/10.1186/s12872-017-0517-2.Search in Google Scholar PubMed PubMed Central
36. Muse, ED, Feldman, DI, Blaha, MJ, Dardari, ZA, Blumenthal, RS, Budoff, MJ, et al.. The association of resistin with cardiovascular disease in the multi-ethnic study of atherosclerosis. Atherosclerosis 2015;239:101–8. https://doi.org/10.1016/j.atherosclerosis.2014.12.044.Search in Google Scholar PubMed PubMed Central
37. Joksić, J, Sopić, M, Spasojević‐Kalimanovska, V, Gojković, T, Zeljković, A, Vekic, J, et al.. Higher circulating resistin protein and PBMCs resistin mRNA levels are associated with increased prevalence of small dense LDL particles in coronary artery disease patients. Clin Exp Pharmacol Physiol 2016;43:22–8. https://doi.org/10.1111/1440-1681.12503.Search in Google Scholar PubMed
38. Li, Y, Wu, QH, Jiao, ML, Fan, XH, Hu, Q, Hao, YH, et al.. Gene-environment interaction between adiponectin gene polymorphisms and environmental factors on the risk of diabetic retinopathy. J Diabetes Investig 2015;6:56–66. https://doi.org/10.1111/jdi.12249.Search in Google Scholar PubMed PubMed Central
39. Steppan, CM, Bailey, ST, Bhat, S, Brown, EJ, Banerjee, RR, Wright, CM, et al.. The hormone resistin links obesity to diabetes. Nature 2001;409:307–12. https://doi.org/10.1038/35053000.Search in Google Scholar PubMed
40. Cho, YM, Youn, BS, Chung, SS, Kim, KW, Lee, HK, Yu, KY, et al.. Common genetic polymorphisms in the promoter of resistin gene are major determinants of plasma resistin concentrations in humans. Diabetologia 2004;47:559–65. https://doi.org/10.1007/s00125-003-1319-x.Search in Google Scholar PubMed
41. Osawa, H, Yamada, K, Onuma, H, Murakami, A, Ochi, M, Kawata, H, et al.. The G/G genotype of a resistin single-nucleotide polymorphism at -420 increases type 2 diabetes mellitus susceptibility by inducing promoter activity through specific binding of Sp1/3. Am J Hum Genet 2004;75:678–86. https://doi.org/10.1086/424761.Search in Google Scholar PubMed PubMed Central
42. Cnop, M, Havel, PJ, Utzschneider, KM, Carr, DB, Sinha, MK, Boyko, EJ, et al.. Relationship of adiponectin to body fat distribution, insulin sensitivity and plasma lipoproteins: evidence for independent roles of age and sex. Diabetologia 2003;46:459–69. https://doi.org/10.1007/s00125-003-1074-z.Search in Google Scholar PubMed
43. Tena-Sempere, M, Pinilla, L, Gonzalez, LC, Dieguez, C, Casanueva, FF, Aguilar, E. Leptin inhibits testosterone secretion from adult rat testis in vitro. J Endocrinol 1999;161:211–8. https://doi.org/10.1677/joe.0.1610211.Search in Google Scholar PubMed
44. Wabitsch, M, Blum, WF, Muche, R, Braun, M, Hube, F, Rascher, W, et al.. Contribution of androgens to the gender difference in leptin production in obese children and adolescents. J Clin Investig 1997;100:808–13. https://doi.org/10.1172/jci119595.Search in Google Scholar PubMed PubMed Central
45. Gabriely, I, Ma, XH, Yang, XM, Atzmon, G, Rajala, MW, Berg, AH, et al.. Removal of visceral fat prevents insulin resistance and glucose intolerance of aging. an adipokine-mediated process? Diabetes 2002;51:2951–8. https://doi.org/10.2337/diabetes.51.10.2951.Search in Google Scholar PubMed
46. Price, JF, Stewart, MCW, Douglas, AF, Murray, GD, Fowkes, GF. Frequency of a low ankle brachial index in the general population by age, sex and deprivation: cross-sectional survey of 28,980 men and women. Eur J Cardiovasc Prev Rehabil 2008;15:370–5. https://doi.org/10.1097/hjr.0b013e3282f8b36a.Search in Google Scholar PubMed
47. Felício, JS, de Melo, FTC, Vieira, GM, de Aquino, VT, de Souza Parente, F, da Silva, WM, et al.. Peripheral arterial disease progression and ankle brachial index: a cohort study with newly diagnosed patients with type 2 diabetes. BMC Cardiovasc Disord 2022;22:294. https://doi.org/10.1186/s12872-022-02722-6.Search in Google Scholar PubMed PubMed Central
48. Morillas, P, Quiles, J, Cordero, A, Guindo, J, Soria, F, Mazon, P, et al.. Impact of clinical and subclinical peripheral arterial disease in mid-term prognosis of patients with acute coronary syndrome. Am J Cardiol 2009;104:1494–8. https://doi.org/10.1016/j.amjcard.2009.07.014.Search in Google Scholar PubMed
49. Zieba, DA, Biernat, W, Szczesna, M, Kirsz, K, Misztal, T. hypothalamic–pituitary and adipose tissue responses to the effect of resistin in sheep: the integration of leptin and resistin signaling involving a suppressor of cytokine signaling 3 and the long form of the leptin receptor. Nutrients 2019;11:2180. https://doi.org/10.3390/nu11092180.Search in Google Scholar PubMed PubMed Central
50. Asterholm, IW, Rutkowski, JM, Fujikawa, T, Cho, YR, Fukuda, M, Tao, C, et al.. Elevated resistin levels induce central leptin resistance and increased atherosclerotic progression in mice. Diabetologia 2014;57:1209–18. https://doi.org/10.1007/s00125-014-3210-3.Search in Google Scholar PubMed PubMed Central
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